Jian Zhao1, Lin Cui2, Jing Sun1, Zhouliang Xie1, Leilei Zhang1, Zhiwei Ding1, Xiaoqiang Quan3. 1. Department of Cardiovascular Surgery, Fuwai Central China Cardiovascular Hospital, People's Hospital of Zhengzhou University, Henan Provincial People's Hospital, Zhengzhou, Henan, China. 2. Department of Cardiovascular Surgery, Fuwai Central China Cardiovascular Hospital, People's Hospital of Zhengzhou University, Henan Provincial People's Hospital, Zhengzhou, Henan, China; Intensive Care Unit, Yellow River Central Hospital, Zhengzhou, Henan, China. 3. Department of Cardiovascular Surgery, Fuwai Central China Cardiovascular Hospital, People's Hospital of Zhengzhou University, Henan Provincial People's Hospital, Zhengzhou, Henan, China. Electronic address: wxslun@163.com.
Abstract
BACKGROUND: Atherosclerosis is a common reason for acute cardio-cerebral vascular diseases. The purpose of this study was to clarify the functional effects of Notoginsenoside R1 (NGR1) on atherosclerosis. METHODS: HUVECS were exposed to oxidized low-density lipoprotein (ox-LDL) in the current study. The proliferation ability of HUVECS was measured by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl-2H-tetrazol-3-ium bromide (MTT). Flow cytometry assays were performed to evaluate the apoptosis of HUVECS. Ox-LDL caused inflammatory response and oxidative stress were assessed by checking pro-inflammatory cytokines and intracellular reactive oxygen species (ROS), malondialdehyde (MDA), and superoxide dismutase (SOD). The regulatory roles of NGR1 in HUVECs were measured by real-time quantitative polymerase chain reaction (RT-qPCR) and western blot assays. The interaction relationship between miR-221-3p and X-inactive specific transcript (XIST) or TNF-receptor-associated factor 6 (TRAF6) was predicted by bioinformatics tools. Dual-luciferase reporter and RNA pull-down assays were used to confirm the interaction relationship. RESULTS: Currently, ox-LDL inhibited proliferation and induced apoptosis, inflammatory response, and oxidative stress in HUVECs, which were alleviated by treatment with NGR1. Importantly, the increase of XIST in ox-LDL-induced HUVECs was abolished by NGR1. In addition, the gain-of-functional experiment suggested that the upregulation of XIST neutralized the protection effects of NGR1 in HUVECs treated with ox-LDL. In addition, miR-221-3p was a target of XIST in HUVECs as confirmed by dual-luciferase reporter and RNA pull-down assays. Furthermore, miR-221-3p interacted with TRAF6, and NGR1 regulated proliferation, apoptosis, inflammatory response, and oxidative stress in HUVECs exposed to ox-LDL by regulation of the XIST/miR-221-3p/TRAF6 axis through Nuclear Factor Kappa B (NF-κB) pathway. CONCLUSION: NGR1 could exert regulatory functions in ox-LDL-induced HUVECS by regulation of XIST/miR-221-3p/TRAF6 axis, which provided valuable insights into the development of potential therapeutic strategy for atherosclerosis.
BACKGROUND:Atherosclerosis is a common reason for acute cardio-cerebral vascular diseases. The purpose of this study was to clarify the functional effects of Notoginsenoside R1 (NGR1) on atherosclerosis. METHODS: HUVECS were exposed to oxidized low-density lipoprotein (ox-LDL) in the current study. The proliferation ability of HUVECS was measured by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl-2H-tetrazol-3-ium bromide (MTT). Flow cytometry assays were performed to evaluate the apoptosis of HUVECS. Ox-LDL caused inflammatory response and oxidative stress were assessed by checking pro-inflammatory cytokines and intracellular reactive oxygen species (ROS), malondialdehyde (MDA), and superoxide dismutase (SOD). The regulatory roles of NGR1 in HUVECs were measured by real-time quantitative polymerase chain reaction (RT-qPCR) and western blot assays. The interaction relationship between miR-221-3p and X-inactive specific transcript (XIST) or TNF-receptor-associated factor 6 (TRAF6) was predicted by bioinformatics tools. Dual-luciferase reporter and RNA pull-down assays were used to confirm the interaction relationship. RESULTS: Currently, ox-LDL inhibited proliferation and induced apoptosis, inflammatory response, and oxidative stress in HUVECs, which were alleviated by treatment with NGR1. Importantly, the increase of XIST in ox-LDL-induced HUVECs was abolished by NGR1. In addition, the gain-of-functional experiment suggested that the upregulation of XIST neutralized the protection effects of NGR1 in HUVECs treated with ox-LDL. In addition, miR-221-3p was a target of XIST in HUVECs as confirmed by dual-luciferase reporter and RNA pull-down assays. Furthermore, miR-221-3p interacted with TRAF6, and NGR1 regulated proliferation, apoptosis, inflammatory response, and oxidative stress in HUVECs exposed to ox-LDL by regulation of the XIST/miR-221-3p/TRAF6 axis through Nuclear Factor Kappa B (NF-κB) pathway. CONCLUSION: NGR1 could exert regulatory functions in ox-LDL-induced HUVECS by regulation of XIST/miR-221-3p/TRAF6 axis, which provided valuable insights into the development of potential therapeutic strategy for atherosclerosis.